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Evaluation De L'activité Antifongique Et Hémolytique Des Extraits Acétate D

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Evaluation De L'activité Antifongique Et Hémolytique Des Extraits Acétate D الشعبية لجمهورية الجزائرية الديمقراطية RÉPUBLIQUE ALGÉRIENNE DÉMOCRATIQUE ET POPULAIRE وزارة التعليم العالي والبحث العلمي MINISTÈRE DE L’ENSEIGNEMENT SUPÉRIEUR ET DE LA RECHERCHE SCIENTIFIQUE Université Frères Mentouri Constantine 1 Département de Biochimie et Biologie Cellulaire et Moléculaire Mémoire présenté en vue de l'obtention du diplôme de MASTER Domaine : Sciences de la Nature et de la Vie Filière : Sciences Biologiques Spécialité : Biochimie Appliquée Thème Evaluation de l’activité antifongique et hémolytique des extraits acétate d’éthyle, n-butanol et aqueux des feuilles d’Abies numidica Soutenu le : 17/07/2019 Par : SENGOUGA Chehinez DENDANI Oumeima Jury d’évaluation : M. KHELIFI Douadi Professeur ENSB Président Mme. BELLIL Inès MCA Univ. Constantine 1 Encadrante Mme MOSBAH Asma MCA Univ. Constantine 1 Examinatrice Année universitaire 2018/2019 “Nothing in biology makes sense except in the light of evolution “ Th. Dobzhansky (1973) Remerciements En préambule à ce mémoire, louange à Dieu le tout miséricordieux pour son guide, son aide dans un parcours acharne envers le savoir scientifique et qui nous a permis de mener à bien ce travail Nous adressons nos sincères remerciements à Mme « BELLIL Ines » maître de conférences (Classe A) à la faculté SNV, Université Frères Mentouri – Constantine 1, d’avoir accepté de nous encadrer, nous la remercions pour sa disponibilité et son aide tout le long de ce modeste travail, qu’elle trouve ici toutes nos gratitudes. Nous vous en sommes très reconnaissantes et en espérant être à la hauteur de votre confiance Nous remercions également la doctorante BENOUCHENNE Djamila pour ses conseils précieux, sa gentillesse et sa disponibilité tout au long de la réalisation de ce mémoire, ainsi que pour sa grande patience, sa modestie, sa générosité. Nous remercions les membres de jury d’avoir bien voulu accepter de juger ce travail. Que Professeur. KHELIFI DOUADI, trouve ici l'expression de nos respectueuses gratitudes et le témoignage de nos profonds remerciements pour avoir accepté de présider ce jury et avant tout pour avoir accepté de nous accueillir dans son laboratoire pour effectuer ce travail. Nous remercions également l’examinatrice de ce travail, MOSBAH ASMA. Maitre DE CONFERENCE A. Nous vous adressons nos sincères remerciements et nos profonds respects pour l’intérêt que vous apportez à ce travail. Nos remercîments s’adressent aussi à tout le personnel du laboratoire de mycologie de Centre de Recherche en Biotechnologie de Constantine pour leur collaboration. Nos remerciements les plus sincères à toutes les personnes qui ont contribué de près ou de loin à l'élaboration de ce mémoire. Dédicace Grace a la volonté divine d’Allah notre dieu tout puissant et bien veillant qui a tracé le chemin de ma vie, j’ai pu réaliser ce modeste travail, que je dédié ! A mes très chers parents : Rachid et Fatima Pour les encouragements, sacrifices, tendresse, amour et soutien durant mes études, je souhaite que j’ai réalisé l’un de vos rêves par ce modeste travail. Puise Dieu vous accorder longue vie pleine de santé et de bonheur. A mon fiancé et mon amour : Fares qui m’encourage toujours à aller plus loin. Merci de m’avoir montré beaucoup de patience durant les moments les plus stressants ainsi que son encouragement A mes chers frères : Aymene, Ahmed Yacine A mes chères sœurs : Rafah, Allae A ma chère tante : Abla et ces filles Norssine, Bailassene A Ma très chère grand-mère A ma binôme et chère amie : Dendeni Oumeima A mes chèreS amie s : Kaouther, Massilia, Nesrine, Sara A tous ma famille A tous mes enseignants depuis le primaire jusqu’à l’université A toute personne ayant contribué de près ou loin à la réalisation de ce travail A toute mes collèges de la promotion de Master II Biochimie appliquée je leur souhaite beaucoup de réussite. CHEHINEZ Dédicace Avant tout, je remercie le BON DIEU le tout miséricordieux Qui m’a donné la patience et la volonté pour arriver à ce stade et réaliser ce travail, que je dédie aux êtres les plus chères à mon cœur : A la meilleure de toutes les mères, source d’amour et d’affection TALEHA RATIBA A la prunelle de mes yeux, l’homme de ma vie mon très cher père MOHAMED Mes chers et tendres parents, merci merci et milles merci pour vos prières, pour tout le soutien que vous me portez depuis mon enfance, pour les sacrifices que vous avez consenti pour mon instruction et mon bien être, pour votre amour infini et votre bienveillance jour et nuit, j’espère que ce modeste travail soit l’exaucement de vos veux tant formulés. Puisse dieu, vous garder à mes côtés le plus longtemps possible. Je vous adore A mon très cher frère : TADJ EDDINE NAWFEL A mes chères sœurs : IMENE, CHAHLA, SIHEM Que dieu nous garde si tendres et aimants les uns envers les autres A mes adorables nièces : MIRNA, SALSSABIL, ASSIL A mon binôme et chère amie : SENGOUGA CHAHINEZ A mes très chères amies : ASMA, MANEL, IBTISSEM, SARA Un grand merci et une forte pensée a tous mes autres amis Et collègues que je ne pourrais citer en entier, merci pour Ces merveilleuses années passées ensembles A tous ceux que j’aime et ceux qui m’aiment OUMEIMA Sommaire Liste des abréviations…………………………………………………………………………….Ⅰ Liste des tableaux…………………………………………………………………………………Ⅱ Liste des figures..…………………………………………………………………………………Ⅲ Résume ……………………………………………………………………………………………Ⅳ Abstract…………………………………………………………………………………………….Ⅴ Ⅵ..…........................................................................................................................................... ملخص Introduction ……………………………………………………………………………………...01 Chapitre 01 : synthèse bibliographique Partie 01 : Abies numidica 1. Historique……………………………………………………………………………………03 2. Distribution géographique…………………………………………………………………03 2.1 Répartition dans le monde et les pays méditerranéennes……………………………03 2.3 Répartition en Algérie …………………………………………………………………04 3. Description botanique………………………………………………………………………05 4.Nomenclature………………………………………………………………………………...06 5.Classification systématique …………………………………………………………………07 6. Usage traditionnelle et thérapeutique………………………………………………………07 Partie 02 : Les Métabolites Secondaires 1. Les composés phénoliques……………………………………………………………...…09 1.1. Les acides phénoliques.. …………………………………………………………….09 1.2. Les flavonoïdes..……………………………………………………………..………10 1.3. Les tannins..………………………………………………………………………….12 1.3.1.les tannins hydrolysables.……………………………………………………..12 1.3.2.les tannins condensé.…………………………………………………………..13 1.4. Les lignines..………………………………………………………………………….13 1.5. Les coumarines..……………………………………………………………………..14 2. Les terpènoides.…………………………………………………………………………..…14 3. Les saponosides.……………………………………………………………………………..15 4. Les alcaloïdes.……………………………………………………………………………….15 5. Activités biologiques de quelques composés phénoliques..………………………………..18 Partie 03 : Présentation de champignon pathogène 1. Les champignons phytopathogènes..………………………………………………….…..19 2. Le Fusarium. .……………………………………………………………………….………19 2.1. Fusarium oxysporium.……………………………………………………………..….19 2.1.1. Classification.. ……………………………………………………………….….20 2.1.2. Morphologie.…………………………………………………………………….21 2.1.3. Cycle de vie..……………………………………………………………………..22 Partie 04 : L’Hémolyse 1. les globules rouges.. ………………………………………………………………………..24 2. la membrane érythrocytaire..……………………………………………………………..24 3. l’hémolyse...…………………………………………………………………………………25 3.1 L’hémolyse physiologique..……………………………………………………………26 3.1.1. L’hémolyse intravasculaire ..………………………………………………….26 3.1.2. L’hémolyse intratissulaire..……………………………………………………26 3.2. L’hémolyse pathologique .……………………………………………………………26 Chapitre 02 : Matériels et Méthodes 1. Préparation de matériel végétal …..……………………………………………………..28 1.1. Echantillonnage ……………………………………………………………………….28 1.2. Extraction et fractionnement des métabolites secondaires…………………………29 1.2.1 Extraction par macération ……………………………………….……………29 1.2.2 Extraction par macération a l’eau………………………………………….…29 1.2.3 Fractionnement de l’extrait méthanolique………………………………..…..29 2. Rendement d’extraction …………………………………………………………………31 3.Screening phytochimique …………………………………………………………………..31 4.Dosage des métabolites secondaires……………………………………………………….33 4.1.Dosage des polyphénols………………………………………………………………..33 4.2.Dosage des flavonoides…………………………………………………………………34 5.Activité antifongique………………………………………………………………………..34 5.1. Préparation de champignon…………………………………………………………..34 5.2. Stérilisation du matériels……………………………………………………………….34 5.3. Préparation des extraits………………………………………………………………..34 5.4. Préparation de milieu de culture………………………………………………………34 5.5. Application des disques mycéliennes………………………………………………….34 6.Evaluation de l’effet hémolytique………………………………………………………………38 6.1. Préparation du suspension érythrocytaire……………………………………………38 6.2. Préparation des extraits………………………………………………………………..38 6.3. Test d’hémolyse…………………………………………………………………………38 6.4. Analyse statistique………………………………………………………………………39 Chapitre 03 : Résultats et Discussions 1.Etude photochimique……………………………………………………………………………40 1.1. Rendement d’extraction………………………………………………………………..40 1.2. Screening photochimique……………………………………………………………….41 1.3. Dosage des métabolites secondaires……………………………………………………43 1.3.1. Dosage des polyphénols…………………………………………………………43 1.3.2. Dosage des flavonoïdes………………………………………………………….45 2.Etudes biologiques……………………………………………………………………………….47 2.1. Activité antifongique…………………………………………………………………….47 2.1.1. Evaluation de la croissance mycélienne……………………………………….47 2.1.1. Pouvoir antifongique …………………………………………………………...52 2.2. Evaluation de l’effet hémolytique
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